Simplified construction and characterization of yeast artificial chromosome libraries.
作者信息
Volinia S, Piva R, Bozza A, Stefani S, Gandini D, del Senno L
机构信息
Instituto di Chimica Biologica, Universit, Ferrara, Italy.
出版信息
Biochem Int. 1992 Jun;27(1):45-53.
PMID:1627178
Abstract
Three yeast artificial chromosome (YAC) libraries were constructed using two human cell lines and the pYAC-RC vector. The main differences from the previously described methods were: i) genomic DNA was digested in low melting point (LMP) agarose blocks with the rare cutting enzyme ClaI; ii) DNA was ligated in melted LMP agarose after agarase treatment; iii) spheroplast regeneration plating was done in calcium alginate thin layer. In addition, a panel of PCR primers was used to identify quickly the presence in the libraries of repetitive and single copy human DNA sequences.
摘要
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